Spectrophotometric method for fast quantification of ascorbic acid and dehydroascorbic acid in simple matrix for...

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Ascorbic acid (AA) and its oxidized form, dehydroascorbic acid (DHAA), are vital antioxidants.
• Accurate quantification of both AA and DHAA is crucial for various biological and industrial applications.
• Existing methods may lack the simplicity, speed, or reliability required for certain applications, such as real-time kinetic monitoring.

Purpose of the Study:

• To develop and validate a simple, rapid, and reliable spectrophotometric method for quantifying both ascorbic acid (AA) and dehydroascorbic acid (DHAA).
• To assess the method's suitability for real-time kinetic measurements in bulk reactors.

Main Methods:

• Spectrophotometric quantification of AA directly or after reduction of DHAA to AA using dithiothreitol.
• Validation of the method in 20mM malate buffer (pH 3.8).
• Assessment of linearity, limits of detection and quantification, fidelity, and accuracy.

Main Results:

• The method demonstrated satisfactory linearity over the 0–6.95mM range.
• Limits of detection and quantification were determined as 0.236mM and 0.467mM, respectively.
• The method showed acceptable fidelity and accuracy, with coefficients of variation below 6% for AA and below 15% for DHAA, and recovery rates of 97–102% for AA and 88–112% for DHAA.

Conclusions:

• A robust and validated spectrophotometric method for quantifying AA and DHAA has been established.
• The developed method is suitable for real-time kinetic studies, offering reliability and accuracy.
• This technique provides a valuable tool for biochemical and industrial analyses requiring precise vitamin C measurements.